Generation of an azide-modified extracellular matrix by adipose-derived stem cells using metabolic glycoengineering

Natural extracellular matrix (ECM) represents an ideal biomaterial for tissue engineering and regenerative medicine approaches. For further functionalization, there is a need for specific addressable functional groups within this biomaterial. Metabolic glycoengineering (MGE) provides a technique to incorporate modified monosaccharide derivatives into the ECM during their assembly, which was shown by us earlier for the production of a modified fibroblast-derived dermal ECM. In this study, adipose-derived stem cells (ASCs) were treated with the azide-modified monosaccharide derivate 1,3,4,6-tetra-O-acetyl-N-azidoacetylgalactosamine (Ac4GalNAz). Toxicity and viability assays after 24 h and 72 h incubation revealed high biocompatibility of Ac4GalNAz in contact with ASCs. The successful incorporation of the functional azide groups into the glycocalyx and the ECM of the ASCs was proven by conjugation with a fluorescent dye via a copper-catalyzed click reaction. Thus, Ac4GalNAz in combination with ASCs was confirmed to achieve an azidemodified ECM as a multifunctional biomaterial for further applications